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Objective The aim is to contribute to the literature by carrying out retrospective analysis of the cases who underwent amniocentesis in our clinic and sharing our relevant experience. Methods A total of 632 cases who underwent amniocentesis in the second trimester were included in our study. The cases’ demographic characteristics, weeks of gestation, amniocentesis indications, procedure-related complications, culture success, cytogenetic results, and the results and indications of the cases found to have chromosomal anomalies were evaluated. Chromosomal anomalies, their complications, culture failure rates and most common amniocentesis indications were reported. The results of the cases with chromosomal anomalies were presented as numerical and structural anomalies. Results The mean age of all cases included in our study was 33.7±6.8 years, and their mean weeks of gestation were 17.5±1.0 weeks. The rate of the cases found to have chromosomal anomaly by amniocentesis was 22.4%, the culture failure rate was 2.1%, and complication rate was 0.5%. The most common amniocentesis indications and the amniocentesis indications in cases found to have chromosomal anomaly were the same, and they were as following respectively: combined test being high risk, triple screening test being high risk and presence of anomaly in the ultrasonography (USG). The rate of detecting chromosomal anomaly was higher in the cases who had multiple indications and underwent amniocentesis. Conclusion Although USG and serum screening tests are used commonly for prenatal diagnosis, invasive tests such as amniocentesis are needed for the final diagnosis in patients with high risk. We concluded that amniocentesis is more important for prenatal diagnosis in cases who are under risk of having multiple chromosomal anomalies in the screening tests.